• Journal of Acupuncture Research
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• 제31권4호
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• pp.33-43
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• 2014

This study was performed to investigate the protective effects of acupuncture on the liver in the oxidative stress caused by cadmium accumulation. Sprague-Dawley male($150{\pm}30g$) rats were stabilized for a week and divided into 5 groups which is normal, control, $LR_3$ acupuncture, $BL_{23}$ acupuncture and sham acupuncture group. For three days experimental groups were received oral doses of cadmium 2 mg/kg twice a day. Acupuncture was given bilaterally at each point 10 times for two weeks. The depth of stimulation was 1 mm at right angles and torsion of acupuncture was produced 2 times per second for 1 minutes. The liver was shipped off and taken weight at the last day of two weeks, and hepatic functions was confirmed through alanine transaminase(ALT) and aspartate amino-transferase(AST). We measured reactive oxygen species of serum, liver and kidney, and compared expression levels of superoxide dismutase(SOD), catalase, glutathione peroxidase(Gpx), nuclear factor erythroid derived 2-related factor 2(Nrf-2), heme oxygenase-1(HO-1), nuclear factor-${\kappa}B$ (NF-${\kappa}B$), cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), Bax and Cytochrome c. $BL_{23}$ acupuncture group significantly increased liver weight and decreased ALT compared to control group. For the oxidative stress, $LR_3$ acupuncture group significantly reduced reactive oxygen species, and $BL_{23}$ acupuncture group significantly reduced reactive oxygen species and inflammation-related protein compared to control group. But $LR_3$ acupuncture group and $BL_{23}$ acupuncture group didn't significantly reduce apoptosis-related protein. Therefore $LR_3$ and $BL_{23}$ acupuncture showed the effects of antioxidant and anti-inflammatory, especially $BL_{23}$ acupuncture was more effective than $LR_3$ acupuncture on the protection of liver in the oxidative stress.

• 한국식품영양과학회지
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• 제43권10호
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• pp.1527-1534
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• 2014

Lactobacillus rhamnosus로 발효시킨 흑마늘 발효물의 항염증 효능을 검증하기 위해 LPS로 염증 유도된 RAW 264.7 cells를 이용하여 관련 인자들을 분석하였다. 100, 200, 400 및 $800{\mu}g/mL$ 농도에서 세포독성은 유발되지 않았으며, 오히려 농도 의존적으로 세포 생존율은 증가하였다. LPS에 의해 염증 유도된 RAW 264.7 cells에서 흑마늘 발효물은 농도 의존적으로 NO와 $PGE_2$의 생성 감소와 염증성 사이토카인인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6의 단백질 생성을 감소시켰다. 또한 iNOS, COX-2, NF-${\kappa}B$ 및 $I{\kappa}B$ 단백질의 발현을 감소시키고 HO-1의 단백질의 발현을 증가시켰다. 이상의 연구 결과를 통해 흑마늘 발효물은 염증에 의한 NF-${\kappa}B$의 활성과 TNF-${\alpha}$, IL-$1{\beta}$와 IL-6의 생성을 억제시키고, iNOS 및 COX-2의 발현을 억제시키는 메커니즘을 통해 염증성 질환의 예방 및 개선 효능을 나타내는 것으로 판단된다.

• 동의신경정신과학회지
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• 제21권2호
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• pp.125-140
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• 2010

Objectives : The purpose of this study is to examine from various angles the protective effect of Gastrodia elata Blume (GEB) against nerve cell death induced by $\beta$-amyloid by using the cell line SH-SY5Y, which is commonly utilized for toxicity testing in nerve cells, and to find out its mechanism of action. Methods : To begin with, as a result of assessing the rate of cell survival by employing MTT reduction assay, the treatment with $\beta$-amyloid at different concentrations caused cytotoxicity, which was inhibited by preprocessing GEB extract. In addition, after $\beta$-amyloid was processed with the cell SH-SY5Y, apoptosis progressed, which was reduced effectively by processing GEB extract. Results : When cytotoxicity was caused by using hydrogen peroxide, a representative ROS, in order to examine the antioxidant effect of GEB, its protective effect was also observed. Apart from ROS, reactive nitrogen species (RNS) are also known to play a crucial role in nerve cell death. The treatment with the NO donor SNAP increased the production of nitric oxide and the expression of iNOS, which was also inhibited by GEB extract. Meanwhile, as an attempt to find out the mechanism of action explaining the antioxidant effect, the intracellular antioxidant enzyme expressions were measured by RT-PCR, which showed that GEB extract increased the expressions of heme oxygenase-1, GAPDH and $\gamma$-glutamate cysteine ligase. Lastly, GEB extract had a protective effect against impaired memory induced by scopolamine in animal models (in vivo). Conclusions : These findings indicate that GEB has a protective effect against the death of cranial nerve cells, suggesting possibilities for the prevention and treatment of AD.

• Molecular & Cellular Toxicology
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• 제2권4호
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• pp.236-243
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• 2006

Microarray analysis of gene expression has become a powerful approach for exploring the biological effects of drugs, particularly at the stage of toxicology and safety assessment. Acetaminophen (APAP) has been known to induce necrosis in liver, but the molecular mechanism involved has not been fully understood. In this study, we investigated gene expression changes of APAP using microarray technology. APAP was orally administered with a single dose of 50 mg/kg or 500 mg/kg into ICR mice and the animals were sacrificed at 6, 24 and 72 h of APAP administration. Serum biochemical markers for liver toxicity were measured to estimate the maximal toxic time and hepatic gene expression was assessed using high-density oligonucleotide microarrays capable of determining the expression profile of >30,000 well-substantiated mouse genes. Significant alterations in gene expression were noted in the liver of APAP-administered mice. The most notable changes in APAP-administered mice were the expression of genes involved in apoptosis, cell cycle, and calcium signaling pathway, cystein metabolism, glutatione metabolism, and MAPK pathway. The majority of the genes upregulated included insulin-like growth factor binding protein 1, heme oxygenase 1, metallothionein 1, S100 calcium binding protein, caspase 4, and P21. The upregulation of apoptosis and cell cycle-related genes were paralleled to response to APAP. Most of the affected gene expressions were returned to control levels after 72 hr. In conclusion, we identified potential hepatotoxicity makers, and these expressions profiling lead to a better understanding of the molecular basis of APAP-induced hapatotoxicity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6651-6661
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• 2015

Breast cancer is a global health concern and is a major cause of death among women. In Oman, it is the most common cancer in women, with an incidence rate of 15.6 per 100,000 Omani females. Various anticancer remedies have been discovered from natural products in the past and the search is continuing for additional examples. Cytotoxic natural compounds may have a major role in cancer therapy either in potentiating the effect of chemotherapy or reducing its harmful effects. Recently, a few studies have reported advantages of using crude camel milk in treating some forms of cancer. However, no adequate data are available on the lyophilised camel's milk responsibility for triggering apoptosis and oxidative stress associated with human breast cancer. The present study aimed to address the role of the lyophilised camel's milk in inducing proliferation repression of BT-474 and HEp-2 cells compared with the non-cancer HCC1937 BL cell line. Lyophilized camel's milk fundamentally repressed BT-474 cells growth and proliferation through the initiation of either the intrinsic and extrinsic apoptotic pathways as indicated by both caspase-3 mRNA and its action level, and induction of death receptors in BT-474 but not the HEp-2 cell line. In addition, lyophilised camel's milk enhanced the expression of oxidative stress markers, heme-oxygenase-1 and reactive oxygen species production in BT-474 cells. Increase in caspase-3 mRNA levels by the lyophilised camel's milk was completely prevented by the actinomycin D, a transcriptional inhibitor. This suggests that lyophilized camel's milk increased newly synthesized RNA. Interestingly,it significantly (p<0.003) repressed the growth of HEp-2 cells and BT-474 cells after treatment for 72 hours while 24 hours treatment repressed BT-474 cells alone. This finding suggests that the lyophilised camel's milk might instigate apoptosis through initiation of an alternative apoptotic pathway.

• 한국작물학회지
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• 제48권2호
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• pp.127-133
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• 2003

The purpose of this study is to investigate the positive or negative effects of 5-aminolevulinic acid(ALA) on the growth of several crops and weeds, by applying a seed soaking treatment, foliar treatment, and application timing, while comparing biological activity between ALA produced by chemical synthesis (Synthetic-ALA) and extracellularly-accumulated ALA by overexpressing the hemeA gene isolated from Bradyrhizobium japonicum(Bio-ALA). Seed soaking treatment of ALA in barley (five cultivars) and wheat (five cultivars) had not shown positive effects at lower concentrations, 0.05 to 0.5 mM as well as negative effects at higher concentrations, 1 to 30 mM. In rice, there also was no positive effect by seed soaking treatment of ALA at lower concentrations, although the rice became damaged by an application of 5 and 10 mM ALA. Growth in barley cultivars, Ganghossalbori, Naehanssalbori, Songhakbori, Saessalbori, and Daehossalbori were increased up to 14%, 19-51 %, 17-64%, 18-23%, and 22-38% by ALA foliar application at lower concentrations, 0.05 to 0.5 mM, respectively. On the other hand, the growth in barley cultivars was inhibited by ALA foliar application at higher concentrations. Barley responded more positively to ALA foliar application than wheat and rice. The growth stimulation caused by ALA seed soaking treatment was less than by ALA foliar treatment. ALA treatment at the 1.5-leaf stage increased growth of barley by 19-58%, while pretreatment to seeds, post-emergence treatment at 3 days after seeding, 3-leaf stages, and 5-leaf stages had not shown positive effects. Thus, the positive effects of ALA on barley were dependent greatly upon the timing of application and its concentration. Monocots weeds were more sensitive to ALA foliar treatment than dicotyledonous weeds. A monocot weed, Setaria viridis L. was the most susceptible plant to ALA while a dicotyledonous weed, Plantago asiatica L. was the most tolerant. No significant difference in biological activity between bio-ALA and synthetic ALA on barley, wheat, rice, and weed, Ixeris dentate tested was observed. Thus, ALA produced by microorganisms would be a potent substance to be used effectively in agricultural production.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1444-1454
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• 2020

Objective: Cold stress induces oxidative damage and impairs energy status of broilers. N-acetylcysteine (NAC) exhibits antioxidant properties and modulates energy metabolism of animals. This study was conducted to investigate the effects of NAC on energy status and antioxidant capacity of heart and liver in the cold-stressed broilers. Methods: The experiment consisted of 4 treatments in a 2×2 factorial arrangement with two diets (basal diet or plus 0.1% NAC) and two ambient temperatures (thermoneutral [conventional ambient temperature] or cold stress [10℃±1℃ during days 15 to 42]). Results: No ascites were seen in cold-stressed broilers. NAC did not attenuate the impaired growth performance of stressed birds. However, NAC decreased plasma asparagine but increased aspartate levels in cold-stressed birds (p<0.05). NAC reduced hepatic adenosine triphosphate (ATP) but elevated adenosine diphosphate contents in unstressed birds (p<0.05). The hepatic ratio of adenosine monophosphate (AMP) to ATP was increased in birds fed NAC (p<0.05). NAC decreased plasma malondialdehyde (MDA) level and cardiac total superoxide dismutase (T-SOD) activity in unstressed birds, but increased hepatic activities of T-SOD, catalase and glutathione peroxidase in stressed birds (p<0.05). NAC down-regulated hepatic AMP-activated protein kinase but up-regulated cardiac heme-oxigenase mRNA expression in stressed birds, and decreased expression of hepatic peroxisome proliferator-activated receptor coactivator-1α as well as hypoxia-inducible factor-1α in liver and heart of birds. Conclusion: Dietary NAC did not affect energy status but enhanced the hepatic antioxidant capacity by increasing the activities of antioxidant enzymes in cold-stressed broilers.

• BMB Reports
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• 제31권6호
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• pp.595-599
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• 1998

To investigate conformational information of a low oxygen affinity recombinant hemoglobin (rHb) containing $96Val{\rightarrow}Trp$ mutation at the ${\alpha}96$ position, we ave produced rHb (${\alpha}96Val{\rightarrow}Phe$) and rHb (${\alpha}96Val{\rightarrow}Tyr$), using the Escherichia coli expression system and site-directed mutagenesis. The oxygen affinity of rHb (${\alpha}96Val{\rightarrow}Phe$) is similar to that of human normal adult hemoglobin (Hb A). However, the oxygen affinity of rHb (${\alpha}96Val{\rightarrow}Tyr$) showed much lower oxygen affinity than Hb A which is similar to that of rHb (${\alpha}96Val{\rightarrow}Tyr$), providing an opportunity as a potential candidate for a hemoglobin-based blood substitute. Both rHb (${\alpha}96Val{\rightarrow}Phe$) and rHb (${\alpha}96Val{\rightarrow}Tyr)$ showed high cooperativity in oxygen binding. IH-NMR spectroscopy shows that both rHb (${\alpha}96Val{\rightarrow}Phe$) and rHb (${\alpha}96Val{\rightarrow}Tyr$) have very similar tertiary structure around the heme pockets and uaternary structure in the ${\alpha}_1/{\beta}_2$ subunit interface ompared to Hb A. The low oxygen affinity of rHb (${\alpha}96Val{\rightarrow}Tyr$) has been suggested to be due to a hydrogen bond caused by an extra hydroxyl group not present in rHb (${\alpha}96Val{\rightarrow}Phe$). However, investigation of the carbonmonoxy form of rHb (${\alpha}96Val{\rightarrow}Phe$) and (${\alpha}96Val{\rightarrow}Try$) in the presence of inositol hexaphosphate at low temperature suggests that low oxygen affinity of (${\alpha}96Val{\rightarrow}Try$) may arise from a mechanism different to that of rHb (${\alpha}96Val{\rightarrow}Trp$).

• 대한한의학방제학회지
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• 제25권1호
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• pp.1-10
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• 2017

Objectives : The fruit of Prunus mume Siebold & Zucc. has been used as an alternative medicine and functional food in Korea and Japan for preventive and therapeutic purposes. However, its molecular actions and mechanism on anti-inflammatory activity have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of the ethanol extract of P. mume fruit (EEPM) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and sought to understand the associated molecular mechanisms. Methods : Cytotoxicity was assessed by an MTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expression of inducible nitric oxide synthase (iNOS) was analyzed by RT-PCR. In addition, expression levels of iNOS, nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein were detected by Western blotting. Results : Our data indicated that EEPM inhibited NO production in LPS-stimulated RAW264.7 cells in a concentration-dependent manner. At the mRNA and protein levels, EEPM suppressed LPS-induced iNOS expression. On the other hand, EEPM markedly enhanced HO-1 expression, which was associated with an induction and nuclear translocation of Nrf2. Moreover, the inhibitory effect of EEPM against LPS‑induced NO production was significantly enhanced by hemin, a HO-1 inducer; however, EEPM's effect on the production of NO was abolished by zinc protoporphyrin IX, a HO-1 inhibitor. Conclusion : The results suggest that EEPM can act as a suppressor agent on NO production through an activation of Nrf2/HO-1 signaling pathway, and may be a promising candidate for the treatment of inflammatory diseases.

• 대한한의학방제학회지
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• 제25권1호
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• pp.51-68
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• 2017

Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.